The nutrient ion concentrations in the interstitial waters of biofilms (BFs) formed on reed and stone surfaces were investigated in the northern and southern basins of Lake Biwa over several years. The following were observed for both types of BF: 1) Concentrations of ammonium, nitrate, nitrite, and phosphate ions were much (hundreds to thousands of times) higher in the BFs than in the surrounding lake water; 2) the concentration of ions, especially nitrate ions, in the BFs changed seasonally, being higher from winter to spring and lower from summer to autumn, synchronizing with the changes in the lake water; 3) dissolved-form N:P ratios were higher in the lake water than BFs; and 4) the bacterial flora of the BFs differed from that of the lake water, with smaller seasonal variations. The present study reveals for the first time that the inside of BFs in a natural environment is rich in nutrient ions and shows similar seasonal changes as the lake water. The BFs in an aquatic environment provide a microenvironment capable of sustaining a specific bacterial flora different from that in the surrounding lake water.Key words: biofilm, nutrient ions, bacterial flora, Lake Biwa Microbial biofilms (BFs) are ubiquitous in aquatic environments and perform important environmental functions, including nutrient recycling (2, 5, 27) and purification of pollutants (8,14,15). Natural BFs are generally composed of a variety of microbes and their secretory products, such as extracellular polymeric substances (EPS). The EPS-containing matrix confers on the constituent microbes certain advantages over their free-living counterparts (16,17). An important advantage is the ability to trap and retain dissolved nutrients through an entrapment mechanism that has been proposed (4, 7, 11) and proven (9) to depend on ion exchange processes.However, few studies have been conducted on the nutrient conditions inside BFs formed in natural environments over a long period of observation. Therefore, the present study was designed to compare over several years the concentrations of various nutrient ions in the interstitial waters of BFs and the surrounding lake water. An additional aim was to examine the relationship between the nutrient conditions and bacterial flora inside the BFs. Moreover, the BFs formed on reed and stone surfaces were compared, to clarify the influences of the substratum surfaces.Here, we report for the first time that the nutrient conditions in a BF and in the surrounding water are significantly different with respect to concentrations of ions, particularly nitrate ions, while exhibiting similar seasonal variations. The relationships among the nutrient conditions, bacterial flora constitution, and the physicochemical properties of the BFs are discussed.
Materials and Methods
Study area and samplingLake Biwa is located in the central part of the Japanese Archipelago (Lat. 35°15' N, Long. 136°05' E). The lake has a surface area of 670 km 2 and a total storage volume of 2.75×10 10 m 3 , consisting of two basins that differ in wa...